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1998•FEMS Microbiology LettersOpen access

Engineering theluxCDABEgenes fromPhotorhabdus luminescensto provide a bioluminescent reporter for constitutive and promoter probe plasmids and mini-Tn5constructs

Michael K. Winson, Simon Swift, Philip J. Hill, Catriona M. Sims, Gottfried Griesmayr, Barrie W. Bycroft, Paul Williams, Gordon S.A.B. Stewart

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Abstract

The luxCDABE operon of Photorhabdus luminescens has been cloned and engineered as an easily mobilisable cassette flanked by sites for commonly used restriction enzymes. Constitutive and promoter probe plasmids utilising the P. luminescens luxCDABE have been constructed using a number of compatible replicons and antibiotic markers. Complementary to these plasmids, a range of promoterless and constitutive luxCDABE mini-Tn5 derivatives has been constructed. The potential of coupling mini-Tn5 luxCDABE promoter probe transposons with automated luminometry and photometry to screen for mutants that exhibit growth phase variation in gene expression is demonstrated.

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The luxCDABE operon of Photorhabdus luminescens has been cloned and engineered as an easily mobilisable cassette flanked by sites for commonly used restriction enzymes. Constitutive and promoter probe plasmids utilising the P. luminescens luxCDABE have been constructed using a number of compatible replicons and antibiotic markers. Complementary to these plasmids, a range of promoterless and constitutive luxCDABE mini-Tn5 derivatives has been constructed. The potential of coupling mini-Tn5 luxCDABE promoter probe transposons with automated luminometry and photometry to screen for mutants that exhibit growth phase variation in gene expression is demonstrated.

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Available abstract

The luxCDABE operon of Photorhabdus luminescens has been cloned and engineered as an easily mobilisable cassette flanked by sites for commonly used restriction enzymes. Constitutive and promoter probe plasmids utilising the P. luminescens luxCDABE have been constructed using a number of compatible replicons and antibiotic markers. Complementary to these plasmids, a range of promoterless and constitutive luxCDABE mini-Tn5 derivatives has been constructed. The potential of coupling mini-Tn5 luxCDABE promoter probe transposons with automated luminometry and photometry to screen for mutants that exhibit growth phase variation in gene expression is demonstrated.

Key concepts: Bioreporter, Photorhabdus luminescens, Biology, Plasmid, Photorhabdus, Gene cassette, Reporter gene, Operon

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Engineering theluxCDABEgenes fromPhotorhabdus luminescensto provide a bioluminescent reporter for constitutive and promoter probe plasmids and mini-Tn5constructs — Research Paper | ScholarLens